Displaying publications 1 - 20 of 35 in total

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  1. Size-related assessment on viability and insulin secretion of caprine islets in vitro
    Vakhshiteh F, Allaudin ZN, Mohd Lila MA, Hani H
    Xenotransplantation, 2013 02 14;20(2):82-8.
    PMID: 23406308 DOI: 10.1111/xen.12023
    BACKGROUND: The successful isolation, purification, and culture of caprine islets has recently been reported. The present study shows arange of size distribution in caprine islet diameter from 50 to 250 μm, in which 80% of the total islet yield was comprised of small islets.

    METHODS: Caprine islets were isolated and purified. Islets were handpicked and the diameter of the islets was recorded using light microscopy. Viablility of the islets was analyzed by confocal microscopy. Insulin secretion assay was carried out and analyzed by ELISA.

    RESULTS: When tested at 48 h after isolation, these small islets were 29.3% more viable compared to the large-sized islets. Large islets showed a high ratio (P 

    Matched MeSH terms: Organ Culture Techniques
  2. Fluorescent cell tracer dye permits real-time assessment of re-epithelialization in a serum-free ex vivo human skin wound assay
    Nasir NAM, Paus R, Ansell DM
    Wound Repair Regen, 2019 01;27(1):126-133.
    PMID: 30575205 DOI: 10.1111/wrr.12688
    Ex vivo wounded human skin organ culture is an invaluable tool for translationally relevant preclinical wound healing research. However, studies incorporating this system are still underutilized within the field because of the low throughput of histological analysis required for downstream assessment. In this study, we use intravital fluorescent dye to lineage trace epidermal cells, demonstrating that wound re-epithelialization of human ex vivo wounds occurs consistent with an extending shield mechanism of collective migration. Moreover, we also report a relatively simple method to investigate global epithelial closure of explants in culture using daily fluorescent dye treatment and en face imaging. This study is the first to quantify healing of ex vivo wounds in a longitudinal manner, providing global assessments for re-epithelialization and tissue contraction. We show that this approach can identify alterations to healing with a known healing promoter. This methodological study highlights the utility of human ex vivo wounds in enhancing our understanding of mechanisms of human skin repair and in evaluating novel therapies to improve healing outcome.
    Matched MeSH terms: Organ Culture Techniques
  3. Des-aspartate angiotensin I (DAA-I) reduces endothelial dysfunction in the aorta of the spontaneously hypertensive rat through inhibition of angiotensin II-induced oxidative stress
    Loh WM, Ling WC, Murugan DD, Lau YS, Achike FI, Vanhoutte PM, et al.
    Vascul. Pharmacol., 2015 Aug;71:151-8.
    PMID: 25869508 DOI: 10.1016/j.vph.2015.03.011
    Des-aspartate angiotensin I (DAA-I), an endogenous nonapeptide, counteracts several effects of angiotensin II on vascular tone. The aim of this study was to investigate the acute protective effect of DAA-I on endothelial function in the spontaneously hypertensive rat (SHR) as well as its effect on angiotensin II-induced contractions and oxidative stress. Aortic rings were incubated with DAA-I (0.1μM) for 30min prior to the assessment of angiotensin II-induced contractions (0.1nM-10μM) in WKY and SHR aortas. Total nitrate and nitrite levels were assessed using a colorimetric method and reactive oxygen species (ROS) were measured by dihydroethidium (DHE) fluorescence and lucigenin-enhanced chemiluminescence. The effect of DAA-I was also assessed against endothelium-dependent and -independent relaxations to acetylcholine and sodium nitroprusside, respectively. Angiotensin II-induced contractions were significantly reduced by DAA-I, losartan and tempol. Incubation with ODQ (soluble guanylyl cyclase inhibitor) and removal of the endothelium prevented the reduction of angiotensin II-induced contractions by DAA-I. Total nitrate and nitrite levels were increased in DAA-I, losartan and tempol treated-SHR tissues while ROS level was reduced by DAA-I and the latter inhibitors. In addition, DAA-I significantly improved the impaired acetylcholine-induced relaxation in SHR aortas whilst sodium nitroprusside-induced endothelium-independent relaxation remained unaffected. The present findings indicate that improvement of endothelial function by DAA-I in the SHR aorta is mediated through endothelium-dependent release of nitric oxide and inhibition of angiotensin II-induced oxidative stress.
    Matched MeSH terms: Organ Culture Techniques
  4. Comparison of chitosan scaffold and chitosan-collagen scaffold: a preliminary study
    Norazril SA, Aminuddin BS, Norhayati MM, Mazlyzam AL, Fauziah O, Ruszymah BH
    Med J Malaysia, 2004 May;59 Suppl B:186-7.
    PMID: 15468880
    Chitosan has similar structure to glycosaminoglycans in the tissue, thus may be a good candidates as tissue engineering scaffold. However, to improve their cell attachment ability, we try to incorporate this natural polymer with collagen by combining it via cross-linking process. In this preliminary study we evaluate the cell attachment ability of chitosan-collagen scaffold versus chitosan scaffold alone. Chitosan and collagen were dissolved in 1% acetic acid and then were frozen for 24 hours before the lyophilizing process. Human skin fibroblasts were seeded into both scaffold and were cultured in F12: DMEM (1:1). Metabolic activity assay were used to evaluate cell attachment ability of scaffold for a period of 1, 3, 7 and 14 days. Scanning electron micrographs shows good cell morphology on chitosan-collagen hybrid scaffold. In conclusion, the incorporation of collagen to chitosan will enhance its cell attachment ability and will be a potential scaffold in tissue engineering.
    Matched MeSH terms: Organ Culture Techniques/methods*
  5. Evaluation of suitable biodegradable scaffolds for engineered bone tissue
    Phang MY, Ng MH, Tan KK, Aminuddin BS, Ruszymah BH, Fauziah O
    Med J Malaysia, 2004 May;59 Suppl B:198-9.
    PMID: 15468886
    Tricalcium phosphate/hydroxyapatite (TCP/HA), hydroxyapatite (HA), chitosan and calcium sulphate (CaSO4) were studied and evaluated for possible bone tissue engineered construct acting as good support for osteogenic cells to proliferate, differentiate, and eventually spread and integrate into the scaffold. Surface morphology visualized by SEM showed that scaffold materials with additional fibrin had more cell densities attached than those without, depicting that the presence of fibrin and collagen fibers were truly a favourite choice of cells to attach. In comparison of various biomaterials used incorporated with fibrin, TCP/HA had the most cluster of cells attached.
    Matched MeSH terms: Organ Culture Techniques/methods*
  6. Collagen as potential cell scaffolds for tissue engineering
    Annuar N, Spier RE
    Med J Malaysia, 2004 May;59 Suppl B:204-5.
    PMID: 15468889
    Selections of collagen available commercially were tested for their biocompatibility as scaffold to promote cell growth in vitro via simple collagen fast test and cultivation of mammalian cells on the selected type of collagen. It was found that collagen type C9791 promotes the highest degree of aggregation as well as cells growth. This preliminary study also indicated potential use of collagen as scaffold in engineered tissue.
    Matched MeSH terms: Organ Culture Techniques/methods
  7. Coral--polyhydroxybutrate composite scaffold for tissue engineering: prefabrication properties
    Al-Salihi KA, Samsudin AR
    Med J Malaysia, 2004 May;59 Suppl B:202-3.
    PMID: 15468888
    In this study the surface properties of two particulate coral and polyhydroxybutrate (PHB) were studied in order to characterize them prior to use in composite production. Coral powder and PHB particle were evaluated using scanning electron microscopy and confocal laser scanning microscopy, to measure surface porosity and pores size. The results showed that coral powder has multiple pleomorphic micropores cross each others give appearance of micro-interconnectivity. Some pore reached to 18 microm with an average porosity of 70%. PHB revealed multiple different size pores extended to the depth, with an average some times reach 25 microm and porosity 45%. These findings demonstrate that both coral and PHB have excellent pores size and porosity that facilitate bone in growth, vascular invasion and bone development. We believe that incorporation of coral powder into PHB will make an excellent composite scaffold for tissue engineering.
    Matched MeSH terms: Organ Culture Techniques/methods
  8. Tissue-engineered bone via seeding bone marrow stem cell derived osteoblasts into coral: a rat model
    Al-Salihi KA
    Med J Malaysia, 2004 May;59 Suppl B:200-1.
    PMID: 15468887
    In the present study, natural coral of porites species was used as scaffold combined with in vitro expanded bone marrow stem cell derived osteoblasts (BMSC-DO), to develop a tissue-engineered bone graft in a rat model. Coral was molded into the shape of rat mandible seeded with 5x10(6) /ml BMSC-DO subsequently implanted subcutaneously in the back of 5 week Sprague dawely rats for 3 months. Coral alone was implanted as a control. The implants were harvest and processed for gross inspection and histological observations. The results showed that newly bone grafts were successfully formed coral seeded with cells group showed smooth highly vascularized like bone tissue. Histological sections revealed mature bone formation and lots of blood vessel, the bone formation occurred in the manner resemble intramembraneous bone formation. This study demonstrates that coral can be use as a suitable scaffold material for delivering bone marrow mesenchymal stem cells in tissue engineering.
    Matched MeSH terms: Organ Culture Techniques/methods*
  9. Design and fabrication of scaffolds for anatomic bone reconstruction
    Hollister SJ, Lin CY, Lin CY, Schek RD, Taboas JM, Flanagan CL, et al.
    Med J Malaysia, 2004 May;59 Suppl B:131-2.
    PMID: 15468853
    Matched MeSH terms: Organ Culture Techniques/methods*
  10. The fundamentals of tissue engineering: new scaffolds
    Di Silvio L, Gurav N, Sambrook R
    Med J Malaysia, 2004 May;59 Suppl B:89-90.
    PMID: 15468832
    The ability to regenerate new bone for skeletal use is a major clinical need. In this study, two novel porous calcium phosphate materials pure HA and biphasic HA/beta-Tricalcium phosphate (HA/beta -TCP) were evaluated as potential scaffolds for cell-seeded bone substitutes using human osteoblast-like cells (HOS) and primary human mesenchymal stem cells (hMSCs). A high rate of proliferation was observed on both scaffolds. A greater increase in alkaline phosphatase (ALP- an indicator of osteoblast differentiation) was observed on HA/beta -TCP compared to HA. This observation indicates that HA/TCP may play a role in inducing osteoblastic differentiation. Although further evaluation is required both materials show potential as innovative synthetic substitutes for tissue engineered scaffolds.
    Matched MeSH terms: Organ Culture Techniques/standards*
  11. Bone marrow mesenchymal stem cells differentiation and proliferation on the surface of coral implant
    Al-Salihi KA, Samsudin AR
    Med J Malaysia, 2004 May;59 Suppl B:45-6.
    PMID: 15468811
    This study was designed to evaluate the ability of natural coral implant to provide an environment for marrow cells to differentiate into osteoblasts and function suitable for mineralized tissue formation. DNA content, alkaline phosptatase (ALP) activity, calcium (Ca) content and mineralized nodules, were measured at day 3, day 7 and day 14, in rat bone marrow stromal cells cultured with coral discs glass discs, while cells alone and coral disc alone were cultured as control. DNA content, ALP activity, Ca content measurements showed no difference between coral, glass and cells groups at 3 day which were higher than control (coral disc alone), but there were higher measurement at day 7 and 14 in the cell cultured on coral than on glass discs, control cells and control coral discs. Mineralized nodules formation (both in area and number) was more predominant on the coral surface than in control groups. These results showed that natural coral implant provided excellent and favorable situation for marrow cell to differentiate to osteoblasts, lead to large amount of mineralized tissue formation on coral surface. This in vitro result could explain the rapid bone bonding of coral in vivo.
    Matched MeSH terms: Organ Culture Techniques*
  12. Strategy for generating tissue-engineered human bone construct
    Tan KK, Aminuddin BS, Tan GH, Sabarul Afian M, Ng MH, Fauziah O, et al.
    Med J Malaysia, 2004 May;59 Suppl B:43-4.
    PMID: 15468810
    The strategy used to generate tissue-engineered bone construct, in view of future clinical application is presented here. Osteoprogenitor cells from periosteum of consenting scoliosis patients were isolated. Growth factors viz TGF-B2, bFGF and IGF-1 were used in concert to increase cell proliferation during in vitro cell expansion. Porous tricalcium phosphate (TCP)-hydroxyapatite (HA) scaffold was used as the scaffold to form 3D bone construct. We found that the addition of growth factors, greatly increased cell growth by 2 to 7 fold. TCP/HA proved to be the ideal scaffold for cell attachment and proliferation. Hence, this model will be further carried out on animal trial.
    Matched MeSH terms: Organ Culture Techniques
  13. Scaffolds in bone tissue engineering
    Nather A
    Med J Malaysia, 2004 May;59 Suppl B:37-8.
    PMID: 15468807
    Matched MeSH terms: Organ Culture Techniques/methods*
  14. Towards engineering of a tracheal equivalent: identification of epithelial precursor cells, differentiation and cocultivation techniques
    Rotter N, Stölzel K, Endres M, Leinhase I, Ziegelaar BW, Sittinger M
    Med J Malaysia, 2004 May;59 Suppl B:35-6.
    PMID: 15468806
    Matched MeSH terms: Organ Culture Techniques
  15. Tissue engineered trachea using sheep nasal septum
    Kojima K
    Med J Malaysia, 2004 May;59 Suppl B:32-3.
    PMID: 15468805
    Matched MeSH terms: Organ Culture Techniques
  16. Autologous human fibrin as the biomaterial for tissue engineering
    Ruszymah BH
    Med J Malaysia, 2004 May;59 Suppl B:30-1.
    PMID: 15468804
    Patient own fibrin may act as the safest, cheapest and immediate available biodegradable scaffold material in clinical 1 tissue engineering. This study investigated the feasibility of using patient own fibrin isolated from whole blood to construct a new human cartilage, skin and bone. Constructed in vitro tissues were implanted on the dorsal part of the nude mice for in vivo maturation. After 8 weeks of implantation, the engineered tissues were removed for histological analysis. Our results demonstrated autologous fibrin has great potential as clinical scaffold material to construct various human tissues.
    Matched MeSH terms: Organ Culture Techniques
  17. Bisnicalaterines B and C, atropisomeric bisindole alkaloids from Hunteria zeylanica, showing vasorelaxant activity
    Hirasawa Y, Hara M, Nugroho AE, Sugai M, Zaima K, Kawahara N, et al.
    J Org Chem, 2010 Jun 18;75(12):4218-23.
    PMID: 20469917 DOI: 10.1021/jo1006762
    Two new bisindole alkaloids, bisnicalaterines B and C (1 and 2) consisting of an eburnane and a corynanthe type of skeletons, were isolated from the bark of Hunteria zeylanica. Their absolute structures were determined by combination of NMR, CD, and computational methods, and each of them was shown to be in an atropisomeric relationship. Bisnicalaterines B and C (1 and 2) showed potent vasorelaxant activity on isolated rat aorta.
    Matched MeSH terms: Organ Culture Techniques
  18. Fulltext Transcription factor CREB3L1 regulates vasopressin gene expression in the rat hypothalamus
    Greenwood M, Bordieri L, Greenwood MP, Rosso Melo M, Colombari DS, Colombari E, et al.
    J Neurosci, 2014 Mar 12;34(11):3810-20.
    PMID: 24623760 DOI: 10.1523/JNEUROSCI.4343-13.2014
    Arginine vasopressin (AVP) is a neurohypophysial hormone regulating hydromineral homeostasis. Here we show that the mRNA encoding cAMP responsive element-binding protein-3 like-1 (CREB3L1), a transcription factor of the CREB/activating transcription factor (ATF) family, increases in expression in parallel with AVP expression in supraoptic nuclei (SONs) and paraventicular nuclei (PVNs) of dehydrated (DH) and salt-loaded (SL) rats, compared with euhydrated (EH) controls. In EH animals, CREB3L1 protein is expressed in glial cells, but only at a low level in SON and PVN neurons, whereas robust upregulation in AVP neurons accompanied DH and SL rats. Concomitantly, CREB3L1 is activated by cleavage, with the N-terminal domain translocating from the Golgi, via the cytosol, to the nucleus. We also show that CREB3L1 mRNA levels correlate with AVP transcription level in SONs and PVNs following sodium depletion, and as a consequence of diurnal rhythm in the suprachiasmatic nucleus. We tested the hypothesis that CREB3L1 activates AVP gene transcription. Both full-length and constitutively active forms of CREB3L1 (CREB3L1CA) induce the expression of rat AVP promoter-luciferase reporter constructs, whereas a dominant-negative mutant reduces expression. Rat AVP promoter deletion constructs revealed that CRE-like and G-box sequences in the region between -170 and -120 bp are important for CREB3L1 actions. Direct binding of CREB3L1 to the AVP promoter was shown by chromatin immunoprecipitation both in vitro and in the SON itself. Injection of a lentiviral vector expressing CREB3L1CA into rat SONs and PVNs resulted in increased AVP biosynthesis. We thus identify CREB3L1 as a regulator of AVP transcription in the rat hypothalamus.
    Matched MeSH terms: Organ Culture Techniques
  19. Fulltext Squamous epitheliotropism of Enterovirus A71 in human epidermis and oral mucosa
    Phyu WK, Ong KC, Kong CK, Alizan AK, Ramanujam TM, Wong KT
    Sci Rep, 2017 03 21;7:45069.
    PMID: 28322333 DOI: 10.1038/srep45069
    Hand-foot-and-mouth disease is a self-limiting paediatric infectious disease commonly caused by Enterovirus A71 (Genus: Enterovirus, Family: Picornaviridae). Typical lesions in and around the hands, feet, oral cavity and other places may rarely be complicated by acute flaccid paralysis and acute encephalomyelitis. Although virus is readily cultured from skin vesicles and oral secretions, the cellular target/s of Enterovirus A71 in human skin and oral mucosa are unknown. In Enterovirus A71-infected human skin and oral mucosa organotypic cultures derived from the prepuce and lip biopsies, focal viral antigens and viral RNA were localized to cytoplasm of epidermal and mucosal squamous cells as early as 2 days post-infection. Viral antigens/RNA were associated with cytoplasmic vacuolation and cellular necrosis. Infected primary prepuce epidermal keratinocyte cultures showed cytopathic effects with concomitant detection of viral antigens from 2 days post-infection. Supernatant and/or tissue homogenates from prepuce skin organotypic cultures and primary prepuce keratinocyte cultures showed viral titres consistent with active viral replication. Our data strongly support Enterovirus A71 squamous epitheliotropism in the human epidermis and oral mucosa, and suggest that these organs are important primary and/or secondary viral replication sites that contribute significantly to oral and cutaneous viral shedding resulting in person-to-person transmission, and viraemia, which could lead to neuroinvasion.
    Matched MeSH terms: Organ Culture Techniques
  20. Bronchodilator effects of Lignosus rhinocerotis extract on rat isolated airways is linked to the blockage of calcium entry
    Lee MK, Lim KH, Millns P, Mohankumar SK, Ng ST, Tan CS, et al.
    Phytomedicine, 2018 Mar 15;42:172-179.
    PMID: 29655683 DOI: 10.1016/j.phymed.2018.03.025
    BACKGROUND: Lignosus rhinocerotis (Cooke) Ryvarden is a popular medicinal mushroom used for centuries in Southeast Asia to treat asthma and chronic cough. The present study aimed to investigate the effect of this mushroom on airways patency.

    MATERIALS AND METHODS: The composition of L. rhinocerotis TM02 cultivar was analyzed. Organ bath experiment was employed to study the bronchodilator effect of Lignosus rhinocerotis cold water extract (CWE) on rat isolated airways. Trachea and bronchus were removed from male Sprague-Dawley rats, cut into rings of 2 mm, pre-contracted with carbachol before adding CWE into the bath in increasing concentrations. To investigate the influence of incubation time, tissues were exposed to intervals of 5, 15 and 30 min between CWE concentrations after pre-contraction with carbachol in subsequent protocol. Next, tissues were pre-incubated with CWE before the addition of different contractile agents, carbachol and 5-hydroxytrptamine (5-HT). The bronchodilator effect of CWE was compared with salmeterol and ipratropium. In order to uncover the mechanism of action of CWE, the role of beta-adrenoceptor, potassium and calcium channels was investigated.

    RESULTS: Composition analysis of TM02 cultivar revealed the presence of β-glucans and derivatives of adenosine. The extract fully relaxed the trachea at 3.75 mg/ml (p 

    Matched MeSH terms: Organ Culture Techniques
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